Hi,

I would like to know why "reduced glutathione" is used to elute the protein? why not oxidised?

Tks=)

HI,
To answer your question, first of all GSH - reduced Gluthathaione is the aboundant form in the cell after the cell was exposed to oxidative stress.
So from what I understand there isn't a oxidized gluththione that is stable.
Here is a link to an explanation about GST
http://www.chemicon.com/company/GST.asp

Here is a link for elution of GST
http://www.upstate.com/misc/protocols.q.prot.e.elution/Elution+of+GST+Fusion+Protein+from+Glutathione+Agarose
Guy

THANKS!=)

Rainydays said:

Hi, I would like to know why "reduced glutathione" is used to elute the protein? why not oxidised? Tks=)

Reduced glutathione is a must for elution and not oxidized because, GST binds to only reduced form. And the Sepharose beads contain reduced glutathione which is competed out by the reduced glutathione we add and thus we get our protein eluted.